Extending the limits of cryo-EM to better understand TTR misfolding and aggregation

扩展冷冻电镜的局限性以更好地了解 TTR 错误折叠和聚集

基本信息

批准号：
10263946
负责人：
Gabriel C Lander
金额：
$ 26.63万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-15 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10263946
关键词：
3-Dimensional Adopted Affect Age Alzheimer&apos s Disease Alzheimer&apos s disease patient Amyloid Amyloid Fibrils Amyloid beta-Protein Amyloidosis Architecture Binding Binding Proteins Biochemical Cardiomyopathies Carrier Proteins Cells Cerebrospinal Fluid Complex Congestive Heart Failure Cryoelectron Microscopy Crystallization Degenerative Disorder Development Diastolic heart failure Disease Disease Progression Dissociation Drug Design Event Funding Future Goals Grant Hepatocyte Imaging technology Individual Kinetics Lead Life Expectancy Ligand Binding Ligands Light Link Methodology Methods Mitotic Molecular Molecular Conformation Molecular Probes Names Pathogenesis Pathologic Pathology Pathway interactions Patients Peptides Pharmaceutical Preparations Plant Roots Population Prealbumin Prevalence Process Property Protein Export Pathway Proteins Protomer Resolution Restrictive Cardiomyopathy Role Serum Structure Therapeutic Therapeutic Intervention Thyroxine Time Tissues Work X-Ray Crystallography age related aggregation pathway amyloidogenesis base combat cytotoxic design fibrillogenesis image processing improved insight monomer network dysfunction non-Native older patient particle prevent protein aggregation protein folding protein misfolding proteostasis proteotoxicity small molecule therapeutic development three dimensional structure

项目摘要

Appl ID: 9981223 Grant Number : 1R21 AG067594-01 PI Name : Lander, Gabriel Grant Title : Extending the limits of cryo-EM to better understand TTR misfolding and aggregation PROJECT SUMMARY The conversion of natively folded proteins into non-functional aggregates is associated with a wide range of age-related degenerative diseases, including Alzheimer’s Disease (AD). Of great concern is the prediction that these diseases will become more prevalent in prevalence as our nation’s life expectancy increases. The development of therapeutics to prevent or reverse the protein misfolding events implicated in degenerative diseases has thus become the focus of many investigative efforts. The protein transthyretin (TTR), a thyroxine and holoretinol carrier protein exported to cerebrospinal fluid (CSF) and serum, is one such protein that demonstrates increased propensity to adopt a non-native fold and form insoluble aggregates with age. This process can occur in the native, wild type form of the TTR protein, and is responsible for wild type TTR amyloidosis (also known as senile systemic amyloidosis), which causes restrictive cardiomyopathy. Notably, there is evidence that TTR interacts with the Aβ peptide, thereby preventing Aβ fibril formation and aggregation. Studies demonstrating that the CSF of AD patients contain substantially lower concentrations of TTR than in the CSF of age-matched non-AD individuals supports a neuroprotective role of TTR. We posit that age-related TTR misfolding and aggregation abolishes the capacity of TTR to prevent Aβ fibril formation and the subsequent onset of AD. In order to better understand how a natively folded wild type TTR protein becomes predisposed to misfolding events, we seek funding to develop structural approaches to study both the native and aberrant forms of TTR. Detailed structural information regarding the destabilization and non-native oligomerization of this protein will profoundly impact our understanding of TTR misfolding and fibrillogenesis, and could lead to the development of more potent TTR stabilizing drugs that could restore neuroprotective properties of TTR in AD patients. The first aim we will push the limits of size and resolution attainable by cryo-electron microscopy to examine the high-resolution three-dimensional structures of destabilized TTR tetramers and compare them to tetramers that are stabilized by small molecule ligands, revealing how the incorporation of misfolded subunits or the improper incorporation of natively folded subunits impact TTR stability at an atomic level. The second aim will define the architecture of TTR aggregates with atomic precision in order to shed light on the assembly pathways and how different oligomeric states differentially contribute to a variety of distinct pathogeneses. Notably, since perturbations of the native folding pathway of wild type proteins are the likely cytotoxic drivers of AD, the findings of this work will likely have far-reaching impact beyond TTR amyloidoses.

应用ID：9981223 赠款编号：1R21 AG067594-01 PI名称：Lander，Gabriel 赠款标题：扩大低温EM的限制以更好地了解TTR错误折叠和一致性项目摘要与年龄相关的变性，包括阿尔茨海默氏病（AD），将天然折叠蛋白转化为非功能性的一致性。预期的发展。 TTR蛋白的天然野生型形式，并导致野生型型淀粉样蛋白（也称为全身性淀粉样蛋白病），这会引起心肌病，这是TTR与Aβ相互作用的Aβ的形成和聚集证明了AD的CSF 患者的TTR浓度低于年龄生的CSF，支持与年龄相关的TTR错误折叠的神经保护作用。，类型的类型会易于折叠事件，以研究TTR的天然和异常形式冷冻电子显微镜检查三维结构化的TTR ttramers，并比较小分子配体稳定的TEM Tetramers y折叠的亚基影响TTR稳定性在第二个原子水平上。病原体由于野生型蛋白的天然途径的扰动是AD淀粉样蛋白以外的AD渗透撞击的可能的细胞毒性驱动因素。